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KMID : 0357119930150020139
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Korean Journal of Immunology
1993 Volume.15 No. 2 p.139 ~ p.146
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Production of Human Monoclonal Antibodies against Tetanus Toxoid using the Epstein-Barr virus transformation
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Yoo Seung-Min
Cho Jeong-Je
Ho Soon-Tae
Ha Youn-Mun
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Abstract
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For production of tetanus toxoid(TT) specific human monoclonal antibodies, anti-TT antibody secreting peripheral B lymphocytes were separated by rosetting with TT-coated SRBC. And, B lymphoblastoid cell line(BLCL) was established by Epstein-Barr Virus(EBV) transformation methods. Stable BLCLs were established after three times of cloning by limiting dilution. To establish the maximal antibody producing condition, several environmental factors were tested. The optimal condition for maximum antibody prodiction was 4 day culture of 1 x 105cells/ml concentration in RPMI 1640 complete culture media supplemented with 20% fetal calf serum, and the maximum concentration of secreted antibody was about 900ng/ml. The antibody production of BLCL was decreased during long-term culture after establishment of cell line, but antibody production was maintained by repeated cloning by limiting dilution.
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KEYWORD
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Epstein Barr virus, Tetanus toxoid, Monoclonal Antibody, BLCL
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